Isolation of Listeria ivanovii from Bulk-Tank Milk of Sheep and Goat Farms-From Clinical Work to Bioinformatics Studies: Prevalence, Association with Milk Quality, Antibiotic Susceptibility, Predictors, Whole Genome Sequence and Phylogenetic Relationships.

Abstract

Simple SummaryAn extensive countrywide study in Greece revealed that isolation of the zoonotic pathogens Listeria monocytogenes and Listeria ivanovii from the milk produced in sheep or goat farms was infrequent: 1.2% of farms sampled. The presence of pigs on the farms, low average relative humidity in the environment and a high number of animals on the farms were found to be associated with the isolations. Detailed assessment of the L. ivanovii strains, for which there is a paucity of information worldwide, revealed that the isolated strains belonged to the L. ivanovii subsp. ivanovii branch. All strains of the branch appeared to be very similar, with the distance between them being small, which suggests that global spread of this clonal branch is a recent evolutionary event or that the branch is characterized by a slow evolutionary rate.A cross-sectional study was performed in 325 sheep and 119 goat dairy farms in Greece. Samples of bulk-tank milk were examined by standard microbiological techniques for Listeria spp. Listeria monocytogenes was isolated from one (0.3%) and Listeria ivanovii from three (0.9%) sheep farms. No associations between the isolation of L. monocytogenes or L. ivanovii and milk quality were found. No resistance to antibiotics was identified. Three variables emerged as significant predictors of isolation of the organism: the presence of pigs, low average relative humidity and a high number of ewes on the farm. The three L. ivanovii isolates were assessed in silico for identification of plasmids, prophages, antibiotic resistance genes, virulence factors, CRISPRs and CAS genes. Phylogenetic analysis using the core genome revealed that the three strains belonged to the L. ivanovii subsp. ivanovii branch and were especially close to the PAM 55 strain. All strains of the branch appeared to be very similar, with the distance between them being small.